Methods of delivering DLL3 antibody drug conjugates

Abstract

Novel modulators, including antibodies and derivatives thereof, and methods of using such modulators to treat proliferative disorders are provided.

The invention claimed is: 
     
       1. A method of delivering a pyrrolobenzodiazepine (PBD) to a DLL3 expressing cancer cell in a subject, the method comprising contacting the cell with a therapeutically effective amount of an antibody drug conjugate (ADC), or a pharmaceutically acceptable salt thereof, wherein the antibody drug conjugate comprises the formula M-[L-D]n, wherein:
 M comprises a humanized anti-DLL3 antibody comprising a light chain variable region set forth as SEQ ID NO: 212 and a heavy chain variable region set forth as SEQ ID NO: 213; 
 L comprises a linker; 
 D comprises a pyrrolobenzodiazepine (PBD) comprising the formula AC: 
 
       
         
           
           
               
               
           
         
         wherein: 
         the dotted lines indicate the optional presence of a double bond, and wherein only one of the dotted lines in a given ring can be a double bond; 
         R 2  is selected from H, OH, ═O, ═CH 2 , CN, R, OR, ═CH—R D , ═C(R D ) 2 , O SO 2  R, CO 2 R, COR, and halo, where R D  is selected from R, CO 2 R, COR, CHO, CO 2 H, and halo; 
         R 6  and R 9  are each independently selected from H, R, OH, OR, SH, SR, NH 2 , NHR, NRR′, NO 2 , Me 3 Sn and halo; 
         R 7  is selected from H, R, OH, OR, SH, SR, NH 2 , NHR, NRR′, NO 2 , Me 3 Sn and halo; 
         R 10  is the linker connected to the anti-DLL3 antibody; 
         Q is selected from O, S and NH; 
         R 11  is either H, or R or, where Q is O, SO 3 M, where M is a metal cation; 
         R and R′ are each independently selected from optionally substituted C 1-12  alkyl, C 3-20  heterocyclyl and C 5-20  aryl groups, and optionally in relation to the group NRR′, R and R′ together with the nitrogen atom to which they are attached form an optionally substituted 4-, 5-, 6- or 7-membered heterocyclic ring; 
         X is selected from O, S, and N(H); 
         R 2″ , R 6″ , R 7″ , R 9″ , and X″ are as defined according to R 2 , R 6 , R 7 , R 9 , and X, respectively; and 
         R″ is a C 3-12  alkylene group, which comprises a chain optionally interrupted by one or more heteroatoms, one or more rings, or both one or more heteroatoms and one or more rings, wherein the optional one or more rings are optionally substituted; and 
         n is an integer from 1 to 20. 
       
     
     
       2. The method of  claim 1 , wherein the cancer cell is a lung cancer cell. 
     
     
       3. The method of  claim 2 , wherein the lung cancer cell is a small cell lung cancer cell. 
     
     
       4. The method of  claim 1 , wherein the cancer cell is a large cell neuroendocrine carcinoma cell. 
     
     
       5. The method of  claim 1 , wherein the cancer cell is a thyroid cancer cell. 
     
     
       6. The method of  claim 1 , wherein the cancer cell is a prostate cancer cell. 
     
     
       7. The method of  claim 1 , wherein the contacting is performed in vitro. 
     
     
       8. The method of  claim 1 , wherein the contacting is performed in vivo. 
     
     
       9. The method of  claim 1 , wherein:
 R 2  is R, wherein R is a C 5-20  aryl group; 
 R 6  and R 9  are H; 
 R 7  is OR, and wherein R is a C 1  alkyl; 
 Q is O, and wherein R 11  is H; and/or 
 X and X″ are O. 
 
     
     
       10. The method of  claim 9 , wherein R 2  is R, wherein R is a C 5-20  aryl group. 
     
     
       11. The method of  claim 9 , wherein R 6  and R 9  are H. 
     
     
       12. The method of  claim 9 , wherein R 7  is OR. 
     
     
       13. The method of  claim 12 , wherein R is a C 1  alkyl. 
     
     
       14. The method of  claim 9 , wherein Q is O. 
     
     
       15. The method of  claim 14 , wherein R 11  is H. 
     
     
       16. The method of  claim 9 , wherein X and X″ are O. 
     
     
       17. A method of determining cytotoxicity of an anti-DLL3 antibody drug conjugate (ADC), or a pharmaceutically acceptable salt thereof, the method comprising the steps of:
 (a) contacting a cancer cell with the antibody drug conjugate, wherein the antibody drug conjugate comprises the formula M-[L-D]n, wherein:
 M comprises a humanized anti-DLL3 antibody comprising a light chain variable region set forth as SEQ ID NO: 212 and a heavy chain variable region set forth as SEQ ID NO: 213; 
 L comprises a linker; 
 D comprises a pyrrolobenzodiazepine (PBD) comprising the formula AC: 
 
 
       
         
           
           
               
               
           
         
         wherein: 
         the dotted lines indicate the optional presence of a double bond, and wherein only one of the dotted lines in a given ring can be a double bond; 
         R 2  is selected from H, OH, ═O, ═CH 2 , CN, R, OR, ═CH—R D , ═C(R D ) 2 , O SO 2  R, CO 2 R, COR, and halo, where R D  is selected from R, CO 2 R, COR, CHO, CO 2 H, and halo; 
         R 6  and R 9  are each independently selected from H, R, OH, OR, SH, SR, NH 2 , NHR, NRR′, NO 2 , Me 3 Sn and halo; 
         R 7  is selected from H, R, OH, OR, SH, SR, NH 2 , NHR, NRR′, NO 2 , Me 3 Sn and halo; 
         R 10  is the linker connected to the anti-DLL3 antibody; 
         Q is selected from O, S and NH; 
         R 11  is either H, or R or, where Q is O, SO 3 M, where M is a metal cation; 
         R and R′ are each independently selected from optionally substituted C 1-12  alkyl, C 3-20  heterocyclyl and C 5-20  aryl groups, and optionally in relation to the group NRR′, R and R′ together with the nitrogen atom to which they are attached form an optionally substituted 4-, 5-, 6- or 7-membered heterocyclic ring; 
         X is selected from O, S, and N(H); 
         R 2″ , R 6″ , R 7″ , R 9″ , and X″ are as defined according to R 2 , R 6 , R 7 , R 9 , and X, respectively; and 
         R″ is a C 3-12  alkylene group, which comprises a chain optionally interrupted by one or more heteroatoms, one or more rings, or both one or more heteroatoms and one or more rings, wherein the optional one or more rings are optionally substituted; and 
         n is an integer from 1 to 20; and 
         (b) determining killing of the cancer cell. 
       
     
     
       18. The method of  claim 17 , wherein the cancer cell is a lung cancer cell. 
     
     
       19. The method of  claim 18 , wherein the lung cancer cell is a small cell lung cancer cell. 
     
     
       20. The method of  claim 17 , wherein the cancer cell is a large cell neuroendocrine carcinoma cell. 
     
     
       21. The method of  claim 17 , wherein the contacting is performed in vitro. 
     
     
       22. The method of  claim 17 , wherein the contacting is performed in vivo. 
     
     
       23. The method of  claim 17 , wherein:
 R 2  is R, wherein R is a C 5-20  aryl group; 
 R 6  and R 9  are H; 
 R 7  is OR, and wherein R is a C 1  alkyl; 
 Q is O, and wherein R 11  is H; and/or 
 X and X″ are O. 
 
     
     
       24. The method of  claim 23 , wherein R 2  is R, wherein R is a C 5-20  aryl group. 
     
     
       25. The method of  claim 23 , wherein R 6  and R 9  are H. 
     
     
       26. The method of  claim 23 , wherein R 7  is OR. 
     
     
       27. The method of  claim 26 , wherein R is a C 1  alkyl. 
     
     
       28. The method of  claim 23 , wherein Q is O. 
     
     
       29. The method of  claim 28 , wherein R 11  is H. 
     
     
       30. The method of  claim 23 , wherein X and X″ are O.